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Jérôme Cornillon, Marie Balsat, Aurélie Cabrespine, Emmanuelle Tavernier-Tardy, Eric Hermet, Aurélien Mulliez, Karine Augeul-Meunier, Denis Guyotat, Jacques-Olivier Bay
Reduced intensity conditioning for allogeneic hematopoietic stem cell transplantation (allo-HSCT) is often proposed for patients with comorbidities. To enhance engraftment and limit graft-versus-host disease (GVHD), antithymoglobulin (ATG) is usually used. However, the dose needed remains unclear unlike myeloablative conditioning. In order to clarify this point, we conducted a retrospective study on patients who received a reduced intensity conditioning allo-HSCT based on a 2-day fludarabine and busulfan treatment with either 1 or 2 days of ATG treatment...
2016: Acta Haematologica
Joanna Kaminska, Weronika Rzepnikowska, Anna Polak, Krzysztof Flis, Piotr Soczewka, Katarzyna Bala, Marzena Sienko, Marcin Grynberg, Pawel Kaliszewski, Agnieszka Urbanek, Kathryn Ayscough, Teresa Zoladek
Human Nedd4 ubiquitin ligase, or its variants, inhibit yeast cell growth by disturbing the actin cytoskeleton organization and dynamics, and lead to an increase in levels of ubiquitinated proteins. In a screen for multicopy suppressors which rescue growth of yeast cells producing Nedd4 ligase with an inactive WW4 domain (Nedd4w4), we identified a fragment of ATG2 gene encoding part of the Atg2 core autophagy protein. Expression of the Atg2-C1 fragment (aa 1074-1447) improved growth, actin cytoskeleton organization, but did not significantly change the levels of ubiquitinated proteins in these cells...
October 2016: International Journal of Biochemistry & Cell Biology
Dah-Shyong Yu, Chia-Lun Wu, Szu-Yuan Ping, Cheng Keng, Kun-Hung Shen
Immunotherapy using bacille Calmette-Guerin (BCG) instillation is the mainstay treatment modality for superficial urothelial cancer (UC) through toll-like receptor (TLR) activation of cognitive immune response. We investigated the roles of TLR7 in the activation of apoptosis in UC cells after BCG treatment. The in vitro cytotoxicity effect of BCG on UC cells was measured by a modified 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl tetrazolium assay. Expressions of TLR7 mRNA and protein in native UC cells prior to and after BCG treatment were analyzed using real-time quantitative polymerase chain reaction and western blot methods...
August 2015: Kaohsiung Journal of Medical Sciences
Y Zhang, R Cai, R Zhou, Y Li, L Liu
Programmed cell death (PCD) has an important role in sculpting organisms during development. However, much remains to be learned about the molecular mechanism of PCD. We found that ectopic expression of tousled-like kinase (tlk) in Drosophila initiated a new type of cell death. Furthermore, the TLK-induced cell death is likely to be independent of the canonical caspase pathway and other known caspase-independent pathways. Genetically, atg2 RNAi could rescue the TLK-induced cell death, and this function of atg2 was likely distinct from its role in autophagy...
January 2016: Cell Death and Differentiation
Sandra Muñoz-Braceras, Rosa Calvo, Ricardo Escalante
Deficient autophagy causes a distinct phenotype in Dictyostelium discoideum, characterized by the formation of multitips at the mound stage. This led us to analyze autophagy in a number of multitipped mutants described previously (tipA(-), tipB(-), tipC(-), and tipD(-)). We found a clear autophagic dysfunction in tipC(-) and tipD(-) while the others showed no defects. tipD codes for a homolog of Atg16, which confirms the role of this protein in Dictyostelium autophagy and validates our approach. The tipC-encoded protein is highly similar to human VPS13A (also known as chorein), whose mutations cause the chorea-acanthocytosis syndrome...
2015: Autophagy
Simon G Pfisterer, Daniela Bakula, Alice Cezanne, Horst Robenek, Tassula Proikas-Cezanne
Macroautophagy (autophagy hereafter) is an evolutionarily highly conserved catabolic process activated by eukaryotes in order to counteract cellular starvation. Autophagy leads to bulk degradation of cytoplasmic content in the lysosomal compartment, thereby clearing the cytoplasm and generating nutrients and energy. Upon autophagy initiation, cytoplasmic material becomes sequestered in newly formed double-membrane vesicles termed 'autophagosomes' that subsequently acquire acidic hydrolases for content destruction...
October 2014: Biochemical Society Transactions
Kwang Deok Shin, Han Nim Lee, Taijoon Chung
Autophagy targets cytoplasmic cargo to a lytic compartment for degradation. Autophagy-related (Atg) proteins, including the transmembrane protein Atg9, are involved in different steps of autophagy in yeast and mammalian cells. Functional classification of core Atg proteins in plants has not been clearly confirmed, partly because of the limited availability of reliable assays for monitoring autophagic flux. By using proUBQ10-GFP-ATG8a as an autophagic marker, we showed that autophagic flux is reduced but not completely compromised in Arabidopsis thaliana atg9 mutants...
May 2014: Molecules and Cells
S P Chantepie, A C Gac, O Reman
In reduced-toxicity conditioning hematopoietic stem cell transplantation, several studies failed to demonstrate the superiority of one conditioning over another. This study described 51 patients (median age of 58 years) allografted with the new FB3-ATG2 conditioning regimen for myeloid (66%) or lymphoid disease (33%). Comorbidity index ≥3 was noted in 23.5% of patients. Toxicities were close to those observed with RIC. One-year cumulative incidence of acute and chronic GVHD was 18.9% and 39.2%. The 2-year-NRM, DFS and OS were 25%, 57...
May 2014: Leukemia Research
Altaf Mohammed, Naveena B Janakiram, Misty Brewer, Rebekah L Ritchie, Anuj Marya, Stan Lightfoot, Vernon E Steele, Chinthalapally V Rao
Epidemiologic studies have shown that diabetes mellitus is associated positively with increased risk of pancreatic ductal adenocarcinoma (PDAC), and recent meta-analysis studies showed that metformin, reduces the risk of pancreatic cancer (PC). We tested the effects of metformin on pancreatic intraepithelial neoplasia (PanIN) and their progression to PDAC in p48Cre/+.LSL-KrasG12D/+ transgenic mice. Mice fed control diet showed 80% and 62% incidence of PDAC in males and females, respectively. Male mice showed 20% and 26%, and female mice showed 7% and 0% PDAC incidence with 1000- and 2000-ppm metformin treatments, respectively...
December 1, 2013: Translational Oncology
Dayana E Salas-Leiva, Alan W Meerow, Javier Francisco-Ortega, Michael Calonje, M Patrick Griffith, Dennis W Stevenson, Kyoko Nakamura
Several individuals of the Caribbean Zamia clade and other cycad genera were used to identify single-copy nuclear genes for phylogeographic and phylogenetic studies in Cycadales. Two strategies were employed to select target loci: (i) a tblastX search of Arabidopsis conserved ortholog sequence (COS) set and (ii) a tblastX search of Arabidopsis-Populus-Vitis-Oryza Shared Single-Copy genes (APVO SSC) against the EST Zamia databases in GenBank. From the first strategy, 30 loci were selected, and from the second, 16 loci...
July 2014: Molecular Ecology Resources
Péter Nagy, Krisztina Hegedűs, Karolina Pircs, Ágnes Varga, Gábor Juhász
The Atg2-Atg18 complex acts in parallel to Atg8 and regulates Atg9 recycling from phagophore assembly site (PAS) during autophagy in yeast. Here we show that in Drosophila, both Atg9 and Atg18 are required for Atg8a puncta formation, unlike Atg2. Selective autophagic degradation of ubiquitinated proteins is mediated by Ref(2)P/p62. The transmembrane protein Atg9 accumulates on refractory to Sigma P (Ref(2)P) aggregates in Atg7, Atg8a and Atg2 mutants. No accumulation of Atg9 is seen on Ref(2)P in cells lacking Atg18 or Vps34 lipid kinase function, while the Atg1 complex subunit FIP200 is recruited...
January 31, 2014: FEBS Letters
Michitaro Shibata, Kazusato Oikawa, Kohki Yoshimoto, Maki Kondo, Shoji Mano, Kenji Yamada, Makoto Hayashi, Wataru Sakamoto, Yoshinori Ohsumi, Mikio Nishimura
The positioning of peroxisomes in a cell is a regulated process that is closely associated with their functions. Using this feature of the peroxisomal positioning as a criterion, we identified three Arabidopsis thaliana mutants (peroxisome unusual positioning1 [peup1], peup2, and peup4) that contain aggregated peroxisomes. We found that the PEUP1, PEUP2, and PEUP4 were identical to Autophagy-related2 (ATG2), ATG18a, and ATG7, respectively, which are involved in the autophagic system. The number of peroxisomes was increased and the peroxisomal proteins were highly accumulated in the peup1 mutant, suggesting that peroxisome degradation by autophagy (pexophagy) is deficient in the peup1 mutant...
December 2013: Plant Cell
Thomas Hackenberg, Trine Juul, Aija Auzina, Sonia Gwizdz, Anna Malolepszy, Katrien Van Der Kelen, Svend Dam, Simon Bressendorff, Andrea Lorentzen, Peter Roepstorff, Kåre Lehmann Nielsen, Jan-Elo Jørgensen, Daniel Hofius, Frank Van Breusegem, Morten Petersen, Stig Uggerhøj Andersen
Programmed cell death often depends on generation of reactive oxygen species, which can be detoxified by antioxidative enzymes, including catalases. We previously isolated catalase-deficient mutants (cat2) in a screen for resistance to hydroxyurea-induced cell death. Here, we identify an Arabidopsis thaliana hydroxyurea-resistant autophagy mutant, atg2, which also shows reduced sensitivity to cell death triggered by the bacterial effector avrRpm1. To test if catalase deficiency likewise affected both hydroxyurea and avrRpm1 sensitivity, we selected mutants with extremely low catalase activities and showed that they carried mutations in a gene that we named NO CATALASE ACTIVITY1 (NCA1)...
November 2013: Plant Cell
Lisa M Farmer, Mauro A Rinaldi, Pierce G Young, Charles H Danan, Sarah E Burkhart, Bonnie Bartel
Peroxisomes house critical metabolic reactions that are essential for seedling development. As seedlings mature, metabolic requirements change, and peroxisomal contents are remodeled. The resident peroxisomal protease LON2 is positioned to degrade obsolete or damaged peroxisomal proteins, but data supporting such a role in plants have remained elusive. Arabidopsis thaliana lon2 mutants display defects in peroxisomal metabolism and matrix protein import but appear to degrade matrix proteins normally. To elucidate LON2 functions, we executed a forward-genetic screen for lon2 suppressors, which revealed multiple mutations in key autophagy genes...
October 2013: Plant Cell
Silvina V Kergaravat, Luis Beltramino, Nidia Garnero, Liliana Trotta, Marta Wagener, Silvia N Fabiano, Maria Isabel Pividori, Silvia R Hernandez
A magneto immunofluorescence assay for the detection of anti-transglutaminase antibodies (ATG2) in celiac disease was developed. The ATG2 were recognized by transglutaminase enzyme immobilized on the magnetic beads and then the immunological reaction was revealed by antibodies labeled with peroxidase. The fluorescent response of the enzymatic reaction with o-phenylenediamine and H2O2 as substrates was correlated with anti-transglutaminase titer, showing EC50 and LOD values of 1:11,600 and 1:74,500 of antibody titers, respectively...
October 10, 2013: Analytica Chimica Acta
Silvina V Kergaravat, Luis Beltramino, Nidia Garnero, Liliana Trotta, Marta Wagener, Maria Isabel Pividori, Silvia R Hernandez
An electrochemical magneto immunosensor for the detection of anti-transglutaminase antibodies (ATG2) in celiac disease was developed. The immunological reaction is performed on magnetic beads (MBs) as a solid support in which the transglutaminase enzyme (TG2) is covalently immobilized (TG2-MB) and then ATG2 were revealed by an antibody labeled with peroxidase. The electrochemical response of the enzymatic reaction with o-phenilendiamine and H₂O₂ as substrates by square wave voltammetry was correlated with the ATG2...
October 15, 2013: Biosensors & Bioelectronics
Rim Marrakchi, Chedly Chouchani, Jeremie Poschmann, Emil Andreev, Mohamed Cherif, Dindial Ramotar
In yeast Saccharomyces cerevisiae, the immunosuppressant rapamycin mimics starvation by inhibiting the kinase Tor1. We recently documented that this treatment triggers a rapid degradation of Sgs1, a helicase involved in several biological processes such as the prevention of genomic instability. Herein, we show that yeast strains deleted for genes ATG2, ATG9, and PEP4, encoding components of the autophagy pathway, prevent rapamycin-induced degradation of Sgs1. We propose that defects in the autophagy pathway prevent degradation of key proteins in the rapamycin response pathway and as a consequence cause resistance to the drug...
June 2013: Biochemistry and Cell Biology, Biochimie et Biologie Cellulaire
Kuninori Suzuki, Manami Akioka, Chika Kondo-Kakuta, Hayashi Yamamoto, Yoshinori Ohsumi
Autophagy is a bulk degradation system mediated by biogenesis of autophagosomes under starvation conditions. In Saccharomyces cerevisiae, a membrane sac called the isolation membrane (IM) is generated from the pre-autophagosomal structure (PAS); ultimately, the IM expands to become a mature autophagosome. Eighteen autophagy-related (Atg) proteins are engaged in autophagosome formation at the PAS. However, the cup-shaped IM was visualized just as a dot by fluorescence microscopy, posing a challenge to further understanding the detailed functions of Atg proteins during IM expansion...
June 1, 2013: Journal of Cell Science
Xiaojun Yang, Xiaomin Zhong, Janos L Tanyi, Jianfeng Shen, Congjian Xu, Peng Gao, Tim M Zheng, Angela DeMichele, Lin Zhang
In human epithelial cancers, the microRNA (miRNA) mir-30d is amplified with high frequency and serves as a critical oncomir by regulating metastasis, apoptosis, proliferation, and differentiation. Autophagy, a degradation pathway for long-lived protein and organelles, regulates the survival and death of many cell types. Increasing evidence suggests that autophagy plays an important function in epithelial tumor initiation and progression. Using a combined bioinformatics approach, gene set enrichment analysis, and miRNA target prediction, we found that mir-30d might regulate multiple genes in the autophagy pathway including BECN1, BNIP3L, ATG12, ATG5, and ATG2...
February 15, 2013: Biochemical and Biophysical Research Communications
Ester Rieter, Fabian Vinke, Daniela Bakula, Eduardo Cebollero, Christian Ungermann, Tassula Proikas-Cezanne, Fulvio Reggiori
Autophagy is a conserved degradative transport pathway. It is characterized by the formation of double-membrane autophagosomes at the phagophore assembly site (PAS). Atg18 is essential for autophagy but also for vacuole homeostasis and probably endosomal functions. This protein is basically a β-propeller, formed by seven WD40 repeats, that contains a conserved FRRG motif that binds to phosphoinositides and promotes Atg18 recruitment to the PAS, endosomes and vacuoles. However, it is unknown how Atg18 association with these organelles is regulated, as the phosphoinositides bound by this protein are present on the surface of all of them...
January 15, 2013: Journal of Cell Science
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